Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.

Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores.

A method for sequencing DNA by using a difluoresceinated primer and laser excitation is described. Dideoxy protocols have been determined that provide sequences for 600 bases starting with base 1 with less than 1% error in a single load. Electrophoresis is at 20 W and the bands are detected 24 cm from the bottom of the loading well with a scanning fluorescence detector. Bands are imaged on a TV...

Plasmid DNA sequencing using highly degenerate oligonucleotides as primers.

Sequencing double-stranded DNA templates has become an efficient procedure (1) being much less time-consuming than the preparation and subsequent sequencing of inserts in phage singlestranded DNA. The ability to use highly degenerate oligonucleotides instead of sequence-specific primers would make this method even more powerful for the rapid sequencing ofDNA that codes for a protein for which a...

Universal Primers for Sequencing DNA Restriction

1.Atcheson, C.L., B. DiDomenico, S. Frackman, R.E. Esposito and R.T. Elder. 1987. Isolation, DNA sequence, and regulation of a meiosis-specific eukaryotic recombination gene. Proc. Natl. Acad. Sci. USA 84:80358039. 2.Cha, J., W. Bishai and S. Chandrasegaran. 1993. New vectors for direct cloning of PCR products [published erratum appears in Gene 1994; 141:149]. Gene 136:369-370. 3.Clark, J.M. 19...

Anchored oligo(dT) primers for automated dye terminator DNA sequencing.

Design strategies and performance of custom DNA sequencing primers.

This study surveyed strategies of sequencing primer selection and evaluated primer performance in automated DNA sequencing. We asked participants to relate their preferred primer design strategies to identify primer characteristics that are considered most important in sequencing primer design. The participants preferred primers of 18-24 nucleotides (nt), 39%-58% G + C, a melting temperature (T...